Canola inbred n00655

ABSTRACT

A novel canola variety designated N00655 and seed, plants and plant parts thereof. Methods for producing a canola plant that comprise crossing canola variety N00655 with another canola plant. Methods for producing a canola plant containing in its genetic material one or more traits introgressed into N00655 through backcross conversion and/or transformation, and to the canola seed, plant and plant part produced thereby. Hybrid canola seed, plant or plant part produced by crossing the canola variety N00655 or a locus conversion of N00655 with another canola variety.

BACKGROUND

A novel rapeseed line designated N00655 is the result of years ofcareful breeding and selection. Since such variety is of high qualityand possesses a relatively low level of erucic acid in the vegetable oilcomponent and a relatively low level of glucosinolate content in themeal component, it can be termed “canola” in accordance with theterminology commonly used by plant scientists.

SUMMARY

Provided a novel Brassica napus line designated N00655. Seed of canolaline N00655, plants of canola line N00655, plant parts of canola lineN00655, and processes for making a canola plant that comprise crossingcanola line N00655 with another Brassica plant are provided. Alsoprovided is N00655 with cytoplasm comprising a gene or genes that causemale sterility. Processes for making a plant containing in its geneticmaterial one or more traits introgressed into N00655 through backcrossconversion and/or transformation, and to the seed, plant and plant partsproduced thereby are provided. A hybrid canola seed, plant or plant partcan be produced by crossing the line N00655 or a locus conversion ofN00655 with another Brassica plant.

Definitions

In the description and examples which follow, a number of terms areused. The following definitions and evaluation criteria are provided.

Anther Fertility. The ability of a plant to produce pollen; measured bypollen production. 1=sterile, 9=all anthers shedding pollen (vs. PollenFormation which is amount of pollen produced).

Anther Arrangement. The general disposition of the anthers in typicalfully opened flowers is observed.

Chlorophyll Content. The typical chlorophyll content of the mature seedsis determined by using methods recommended by the Western CanadaCanola/Rapeseed Recommending Committee (WCC/RRC). 1=low (less than 8ppm), 2=medium (8 to 15 ppm), 3=high (greater than 15 ppm). Also,chlorophyll could be analyzed using NIR (Near Infrared) spectroscopy aslong as the instrument is calibrated according to the manufacturer'sspecifications.

CMS. Abbreviation for cytoplasmic male sterility.

Cotyledon. A cotyledon is a part of the embryo within the seed of aplant; it is also referred to as a seed leaf. Upon germination, thecotyledon may become the embryonic first leaf of a seedling.

Cotyledon Length. The distance between the indentation at the top of thecotyledon and the point where the width of the petiole is approximately4 mm.

Cotyledon Width. The width at the widest point of the cotyledon when theplant is at the two to three-leaf stage of development. 3=narrow,5=medium, 7=wide.

CV %: Abbreviation for coefficient of variation.

Cytoplasmic Conversion. A plant that has been developed by transferringthe cytoplasm of a plant to a variety of interest. This can be donethrough crossing the variety of interest to a plant that has the desiredcytoplasm and backcrossing to the variety of interest. The cytoplasmwill be transferred through the female parent. The result would be thegenome of the variety of interest with the cytoplasm of another plant,generally the cytoplasm from the other plant will confer male sterility.

Disease Resistance: Resistance to various diseases is evaluated and isexpressed on a scale of 0=not tested, 1=resistant, 3=moderatelyresistant, 5=moderately susceptible, 7=susceptible, and 9=highlysusceptible.

Erucic Acid Content: The percentage of the fatty acids in the form ofC22:1.as determined by one of the methods recommended by the WCC/RRC,being AOCS Official Method Ce 2-66 Preparation of Methyl esters ofLong-Chain Fatty Acids or AOCS Official Method Ce 1-66 Fatty AcidComposition by Gas Chromatography.

F1 Progeny. A first generation progeny plant produced by crossing aplant of canola variety S00616BI with a plant of another canola plant.

Fatty Acid Content: The typical percentages by weight of fatty acidspresent in the endogenously formed oil of the mature whole dried seedsare determined. During such determination the seeds are crushed and areextracted as fatty acid methyl esters following reaction with methanoland sodium methoxide. Next the resulting ester is analyzed for fattyacid content by gas liquid chromatography using a capillary column whichallows separation on the basis of the degree of unsaturation and fattyacid chain length.

Flower Bud Location. A determination is made whether typical buds aredisposed above or below the most recently opened flowers.

Flower Date 50%. (Same as Time to Flowering) The number of days fromplanting until 50% of the plants in a planted area have at least oneopen flower.

Flower Petal Coloration. The coloration of open exposed petals on thefirst day of flowering is observed.

Frost Tolerance (Spring Type Only). The ability of young plants towithstand late spring frosts at a typical growing area is evaluated andis expressed on a scale of 1 (poor) to 5 (excellent).

Gene Silencing. The interruption or suppression of the expression of agene at the level of transcription or translation.

Genotype. Refers to the genetic constitution of a cell or organism.

Glucosinolate Content. The total glucosinolates of seed at 8.5%moisture, as measured by AOCS Official Method AK-1-92 (determination ofglucosinolates content in rapeseed—colza by HPLC), is expressed asmicromoles per gram of defatted, oil-free meal. Capillary gaschromatography of the trimethylsityl derivatives of extracted andpurified desulfoglucosinolates with optimization to obtain optimumindole glucosinolate detection is described in “Procedures of theWestern Canada Canola/Rapeseed Recommending Committee Incorporated forthe Evaluation and Recommendation for Registration of Canola/RapeseedCandidate Cultivars in Western Canada”. Also, glucosinolates could beanalyzed using NIR (Near Infrared) spectroscopy as long as theinstrument is calibrated according to the manufacturer's specifications.

Grain. Seed produced by the plant or a self or sib of the plant that isintended for food or feed use.

Green Seed. The number of seeds that are distinctly green throughout asdefined by the Canadian Grain Commission. Expressed as a percentage ofseeds tested.

Herbicide Resistance: Resistance to various herbicides when applied atstandard recommended application rates is expressed on a scale of 1(resistant), 2 (tolerant), or 3 (susceptible).

Leaf Anthocyanin Coloration. The presence or absence of leaf anthocyanincoloration, and the degree thereof if present, are observed when theplant has reached the 9- to 11-leaf stage.

Leaf Attachment to Stem. The presence or absence of clasping where theleaf attaches to the stem, and when present the degree thereof, areobserved.

Leaf Attitude. The disposition of typical leaves with respect to thepetiole is observed when at least 6 leaves of the plant are formed.

Leaf Color. The leaf blade coloration is observed when at least sixleaves of the plant are completely developed.

Leaf Glaucosity or Waxiness. The presence or absence of a fine whitishpowdery coating on the surface of the leaves, and the degree thereofwhen present, are observed.

Leaf Length. The length of the leaf blades and petioles are observedwhen at least six leaves of the plant are completely developed.

Leaf Lobes. The fully developed upper stem leaves are observed for thepresence or absence of leaf lobes when at least 6 leaves of the plantare completely developed.

Leaf Margin Indentation. A rating of the depth of the indentations alongthe upper third of the margin of the largest leaf. 1=absent or very weak(very shallow), 3=weak (shallow), 5=medium, 7=strong (deep), 9=verystrong (very deep).

Leaf Margin Hairiness. The leaf margins of the first leaf are observedfor the presence or absence of pubescence, and the degree thereof, whenthe plant is at the two leaf-stage.

Leaf Margin Shape. A visual rating of the indentations along the upperthird of the margin of the largest leaf. 1=undulating, 2=rounded,3=sharp.

Leaf Surface or Texture. The leaf surface is observed for the presenceor absence of wrinkles when at least six leaves of the plant arecompletely developed.

Leaf Tip Reflexion. The presence or absence of bending of typical leaftips and the degree thereof, if present, are observed at the six toeleven leaf-stage.

Leaf Upper Side Hairiness. The upper surfaces of the leaves are observedfor the presence or absence of hairiness, and the degree thereof ifpresent, when at least six leaves of the plant are formed.

Leaf Width. The width of the leaf blades is observed when at least sixleaves of the plant are completely developed.

Locus. A specific location on a chromosome.

Locus Conversion. A locus conversion refers to plants within a varietythat have been modified in a manner that retains the overall genetics ofthe variety and further comprises one or more loci with a specificdesired trait, such as male sterility, insect, disease or herbicideresistance. Examples of single locus conversions include mutant genes,transgenes and native traits finely mapped to a single locus. One ormore locus conversion traits may be introduced into a single canolavariety.

Lodging Resistance. Resistance to lodging at maturity is observed. 1=nottested, 3=poor, 5=fair, 7=good, 9=excellent.

LSD. Abbreviation for least significant difference.

Maturity or Time to Maturity. The number of days from planting tomaturity is observed, with maturity being defined as the plant stagewhen pods with seed change color, occurring from green to brown orblack, on the bottom third of the pod-bearing area of the main stem.

NMS. Abbreviation for nuclear male sterility.

Number of Leaf Lobes. The frequency of leaf lobes, when present, isobserved when at least six leaves of the plant are completely developed.

Oil Content: The typical percentage by weight oil present in the maturewhole dried seeds is determined by ISO 10565:1993 Oilseeds Simultaneousdetermination of oil and water—Pulsed NMR method. Also, oil could beanalyzed using NIR (Near Infrared) spectroscopy as long as theinstrument is calibrated according to the manufacturer's specifications,reference AOCS Procedure Am 1-92 Determination of Oil, Moisture andVolatile Matter, and Protein by Near-Infrared Reflectance.

Pedicel Length. The typical length of the silique stem when mature isobserved. 3=short, 5=medium, 7=long.

Petal Length. The lengths of typical petals of fully opened flowers areobserved. 3=short, 5=medium, 7=long.

Petal Spacing. Flower petal spacing is the measurement of how the petalsare arranged in the flower. The petals should be observed when they arefully open, but not wilted. Spacing is rated on a scale of 1-9, 1 beingthin petals with a large space between them, and 9 being large leavedwith a strong overlap.

Petal Width. The widths of typical petals of fully opened flowers areobserved. 3=short, 5=medium, 7=long.

Petiole Length. The length of the petioles is observed, in a lineforming lobed leaves, when at least six leaves of the plant arecompletely developed. 3=short, 5=medium, 7=long.

Plant. As used herein, the term “plant” includes reference to animmature or mature whole plant, including a plant that has beenemasculated or from which seed or grain has been removed. Seed or embryothat will produce the plant is also considered to be the plant.

Plant Part. As used herein, the term “plant part” includes leaves,stems, roots, seed, grain, embryo, pollen, ovules, flowers, husks,stalks, root tips, anthers, pericarp, tissue, cells and the like.

Plant Height. The overall plant height at the end of flowering isobserved. 3=short, 5=medium, 7=tall.

Platform indicates the variety with the base genetics and the varietywith the base genetics comprising locus conversion(s). There can be aplatform for the inbred canola variety and the hybrid canola variety.

Ploidy. This refers to the number of chromosomes exhibited by the line,for example diploid or tetraploid.

Pod Anthocyanin Coloration. The presence or absence at maturity ofsilique anthocyanin coloration, and the degree thereof if present, areobserved.

Pod (Silique) Beak Length. The typical length of the silique beak whenmature is observed. 3=short, 5=medium, 7=long.

Pod Habit. The typical manner in which the siliques are borne on theplant at maturity is observed.

Pod (Silique) Length. The typical silique length is observed. 1=short(less than 7 cm), 5=medium (7 to 10 cm), 9=long (greater than 10 cm).

Pod (Silique) Attitude or Angle. A visual rating of the angle joiningthe pedicel to the pod at maturity. 1=erect, 3=semi-erect, 5=horizontal,7=semi-drooping, 9=drooping.

Pod Type. The overall configuration of the silique is observed.

Pod (Silique) Width. The typical pod width when mature is observed.3=narrow (3 mm), 5=medium (4 mm), 7=wide (5 mm).

Pollen Formation. The relative level of pollen formation is observed atthe time of dehiscence.

Protein Content: The typical percentage by weight of protein in the oilfree meal of the mature whole dried seeds is determined by AOCS OfficialMethod Ba 4e-93 Combustion Method for the Determination of CrudeProtein. Also, protein could be analyzed using NIR (Near Infrared)spectroscopy as long as the instrument is calibrated according to themanufacturer's specifications, reference AOCS Procedure Am 1-92Determination of Oil, Moisture and Volatile Matter, and Protein byNear-Infrared Reflectance.

Resistance. The ability of a plant to withstand exposure to an insect,disease, herbicide, or other condition. A resistant plant variety orhybrid will have a level of resistance higher than a comparablewild-type variety or hybrid. “Tolerance” is a term commonly used incrops affected by Sclerotinia, and is used to describe an improved levelof field resistance.

Root Anthocyanin Coloration. The presence or absence of anthocyanincoloration in the skin at the top of the root is observed when the planthas reached at least the six-leaf stage.

Root Anthocyanin Expression. When anthocyanin coloration is present inskin at the top of the root, it further is observed for the exhibitionof a reddish or bluish cast within such coloration when the plant hasreached at least the six-leaf stage.

Root Anthocyanin Streaking. When anthocyanin coloration is present inthe skin at the top of the root, it further is observed for the presenceor absence of streaking within such coloration when the plant hasreached at least the six-leaf stage.

Root Chlorophyll Coloration. The presence or absence of chlorophyllcoloration in the skin at the top of the root is observed when the planthas reached at least the six-leaf stage.

Root Coloration Below Ground. The coloration of the root skin belowground is observed when the plant has reached at least the six-leafstage.

Root Depth in Soil. The typical root depth is observed when the planthas reached at least the six-leaf stage.

Root Flesh Coloration. The internal coloration of the root flesh isobserved when the plant has reached at least the six-leaf stage.

SE. Abbreviation for standard error.

Seedling Growth Habit. The growth habit of young seedlings is observedfor the presence of a weak or strong rosette character. 1=weak rosette,9=strong rosette.

Seeds Per Pod. The average number of seeds per pod is observed.

Seed Coat Color. The seed coat color of typical mature seeds isobserved. 1=black, 2=brown, 3=tan, 4=yellow, 5=mixed, 6=other.

Seed Coat Mucilage. The presence or absence of mucilage on the seed coatis determined and is expressed on a scale of 1 (absent) to 9 (present).During such determination a petri dish is filled to a depth of 0.3 cm.with water provided at room temperature. Seeds are added to the petridish and are immersed in water where they are allowed to stand for fiveminutes. The contents of the petri dish containing the immersed seedsare then examined under a stereo microscope equipped with transmittedlight. The presence of mucilage and the level thereof is observed as theintensity of a halo surrounding each seed.

Seed Size or Seed Weight. The weight in grams of 1,000 typical seeds isdetermined at maturity while such seeds exhibit a moisture content ofapproximately 5 to 6 percent by weight.

Shatter Resistance. Resistance to silique shattering is observed at seedmaturity. 1=not tested, 3=poor, 5=fair, 7=good, 9=does not shatter.

SI. Abbreviation for self-incompatible.

Speed of Root Formation. The typical speed of root formation is observedwhen the plant has reached the four to eleven-leaf stage.

SSFS. Abbreviation for Sclerotinia sclerotiorum Field Severity score, arating based on both percentage infection and disease severity.

Stem Anthocyanin Intensity. The presence or absence of leaf anthocyanincoloration and the intensity thereof, if present, are observed when theplant has reached the nine to eleven-leaf stage. 1=absent or very weak,3=weak, 5=medium, 7=strong, 9=very strong.

Stem Lodging at Maturity. A visual rating of a plant's ability to resiststem lodging at maturity. 1=very weak (lodged), 9=very strong (erect).

Time to Flowering. A determination is made of the number of days when atleast 50 percent of the plants have one or more open buds on a terminalraceme in the year of sowing.

Seasonal Type. This refers to whether the new line is considered to beprimarily a Spring or Winter type of canola.

Variety. A canola line and minor genetic modifications thereof thatretain the overall genetics of the line including but not limited to alocus conversion, a cytoplasm conversion, a mutation, or a somoclonalvariant.

Winter Survival (Winter Type Only). The ability to withstand wintertemperatures at a typical growing area is evaluated and is expressed ona scale of 1 (poor) to 5 (excellent).

DETAILED DESCRIPTION

Field crops are bred through techniques that take advantage of theplant's method of pollination. A plant is self-pollinated if pollen fromone flower is transferred to the same or another flower of the sameplant or a genetically identical plant. A plant is sib-pollinated whenindividuals within the same family or line are used for pollination. Aplant is cross-pollinated if the pollen comes from a flower on agenetically different plant from a different family or line. The term“cross-pollination” used herein does not include self-pollination orsib-pollination.

The breeder often initially selects and crosses two or more parentallines, followed by repeated selfing and selection, thereby producingmany unique genetic combinations. In each cycle of evaluation, the plantbreeder selects the germplasm to advance to the next generation. Thisgermplasm is grown under chosen geographical, climatic, and soilconditions, and further selections are then made. The unpredictabilityof genetic combinations commonly results in the expenditure of largeeffort to develop a new and superior canola variety.

Canola breeding programs utilize techniques such as mass and recurrentselection, backcrossing, pedigree breeding and haploidy.

Recurrent selection is used to improve populations of either self- orcross-pollinating Brassica. Through recurrent selection, a geneticallyvariable population of heterozygous individuals is created byintercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, and/or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued. Variousrecurrent selection techniques are used to improve quantitativelyinherited traits controlled by numerous genes.

Breeding programs use backcross breeding to transfer genes for a simplyinherited, highly heritable trait into another line that serves as therecurrent parent. The source of the trait to be transferred is calledthe donor parent. After the initial cross, individual plants possessingthe desired trait of the donor parent are selected and are crossed(backcrossed) to the recurrent parent for several generations. Theresulting plant is expected to have the attributes of the recurrentparent and the desirable trait transferred from the donor parent. Thisapproach has been used for breeding disease resistant phenotypes of manyplant species, and has been used to transfer low erucic acid and lowglucosinolate content into lines and breeding populations of Brassica.

Pedigree breeding and recurrent selection breeding methods are used todevelop varieties from breeding populations. Pedigree breeding startswith the crossing of two genotypes, each of which may have one or moredesirable characteristics that is lacking in the other or whichcomplements the other. If the two original parents do not provide all ofthe desired characteristics, other sources can be included in thebreeding population. In the pedigree method, superior plants are selfedand selected in successive generations. In the succeeding generationsthe heterozygous condition gives way to homogeneous lines as a result ofself-pollination and selection. Typically, in the pedigree method ofbreeding, five or more generations of selfing and selection arepracticed: F₁ to F₂; F₂ to F₃; F₃ to F₄; F₄ to F₅, etc. For example, twoparents that are believed to possess favorable complementary traits arecrossed to produce an F₁. An F₂ population is produced by selfing one orseveral F₁'s or by intercrossing two F₁'s (i.e., sib mating). Selectionof the best individuals may begin in the F₂ population, and beginning inthe F₃ the best individuals in the best families are selected.Replicated testing of families can begin in the F₄ generation to improvethe effectiveness of selection for traits with low heritability. At anadvanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines commonly are tested forpotential release as new cultivars. Backcrossing may be used inconjunction with pedigree breeding; for example, a combination ofbackcrossing and pedigree breeding with recurrent selection has beenused to incorporate blackleg resistance into certain cultivars ofBrassica napus.

Plants that have been self-pollinated and selected for type for manygenerations become homozygous at almost all gene loci and produce auniform population of true breeding progeny. If desired, double-haploidmethods can also be used to extract homogeneous lines from inbredN00655. A cross between two different homozygous lines produces auniform population of hybrid plants that may be heterozygous for manygene loci. A cross of two plants each heterozygous at a number of geneloci will produce a population of hybrid plants that differ geneticallyand will not be uniform.

The choice of breeding or selection methods depends on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of cultivar used commercially, such as F₁ hybrid variety or openpollinated variety. A true breeding homozygous line can also be used asa parental line (inbred line) in a commercial hybrid. If the line isbeing developed as an inbred for use in a hybrid, an appropriatepollination control system should be incorporated in the line.Suitability of an inbred line in a hybrid combination will depend uponthe combining ability (general combining ability or specific combiningability) of the inbred.

Various breeding procedures can be utilized with these breeding andselection methods and inbred N00655. The single-seed descent procedurein the strict sense refers to planting a segregating population,harvesting a sample of one seed per plant, and using the one-seed sampleto plant the next generation. When the population has been advanced fromthe F₂ to the desired level of inbreeding, the plants from which linesare derived will each trace to different F₂ individuals. The number ofplants in a population declines each generation due to failure of someseeds to germinate or some plants to produce at least one seed. As aresult, not all of the F₂ plants originally sampled in the populationwill be represented by a progeny when generation advance is completed.

In a multiple-seed procedure, one or more pods from each plant in apopulation are threshed together to form a bulk. Part of the bulk isused to plant the next generation and part is put in reserve. Theprocedure has been referred to as modified single-seed descent or thepod-bulk technique. It is considerably faster to thresh pods with amachine than to remove one seed from each by hand for the single-seedprocedure. The multiple-seed procedure also makes it possible to plantthe same number of seeds of a population each generation of inbreeding.Enough seeds are harvested to make up for those plants that did notgerminate or produce seed. If desired, doubled-haploid methods can beused to extract homogeneous lines.

Molecular markers, including techniques such as Isozyme Electrophoresis,Restriction Fragment Length Polymorphisms (RFLP), random amplifiedpolymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP),inter-simple sequence repeats (ISSRs), sequence characterized regions(SCARs), sequence tag sites (STSs), cleaved amplified polymorphicsequences (CAPS), microsatellites, simple sequence repeats (SSRs),expressed sequence tags (ESTs), single nucleotide polymorphisms (SNPs),and diversity arrays technology (DArT), sequencing, and the like may beused in plant breeding methods using N00655. One use of molecularmarkers is Quantitative Trait Loci (QTL) mapping. QTL mapping is the useof markers which are known to be closely linked to alleles that havemeasurable effects on a quantitative trait. Selection in the breedingprocess is based upon the accumulation of markers linked to the positiveeffecting alleles and/or the elimination of the markers linked to thenegative effecting alleles in the plant's genome.

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select for the genome of the recurrent parent and against themarkers of the donor parent. Using this procedure can minimize theamount of genome from the donor parent that remains in the selectedplants. It can also be used to reduce the number of crosses back to therecurrent parent needed in a backcrossing program. The use of molecularmarkers in the selection process is often called Genetic Marker EnhancedSelection or Marker Assisted Selection (MAS).

Methods of isolating nucleic acids from N00655 and methods forperforming genetic marker profiles using SNP and SSR polymorphisms areprovided. SNPs are genetic markers based on a polymorphism in a singlenucleotide. A marker system based on SNPs can be highly informative inlinkage analysis relative to other marker systems in that multiplealleles may be present.

A method comprising isolating nucleic acids, such as DNA, from a plant,a plant part, plant cell or a seed of the canola varieties disclosedherein is provided. The method can include mechanical, electrical and/orchemical disruption of the plant, plant part, plant cell or seed,contacting the disrupted plant, plant part, plant cell or seed with abuffer or solvent, to produce a solution or suspension comprisingnucleic acids, optionally contacting the nucleic acids with aprecipitating agent to precipitate the nucleic acids, optionallyextracting the nucleic acids, and optionally separating the nucleicacids such as by centrifugation or by binding to beads or a column, withsubsequent elution, or a combination thereof. If DNA is being isolated,an RNase can be included in one or more of the method steps. The nucleicacids isolated can comprise all or substantially all of the genomic DNAsequence, all or substantially all of the chromosomal DNA sequence orall or substantially all of the coding sequences (cDNA) of the plant,plant part, or plant cell from which they were isolated. The nucleicacids isolated can comprise all, substantially all, or essentially allof the genetic complement of the plant. The nucleic acids isolated cancomprise a genetic complement of the canola variety. The amount and typeof nucleic acids isolated may be sufficient to permit whole genomesequencing of the plant from which they were isolated or chromosomalmarker analysis of the plant from which they were isolated.

The methods can be used to produce nucleic acids from the plant, plantpart, seed or cell, which nucleic acids can be, for example, analyzed toproduce data. The data can be recorded. The nucleic acids from thedisrupted cell, the disrupted plant, plant part, plant cell or seed orthe nucleic acids following isolation or separation can be contactedwith primers and nucleotide bases, and/or a polymerase to facilitate PCRsequencing or marker analysis of the nucleic acids. In some examples,the nucleic acids produced can be sequenced or contacted with markers toproduce a genetic profile, a molecular profile, a marker profile, ahaplotype, or any combination thereof. In some examples, the geneticprofile or nucleotide sequence is recorded on a computer readablemedium. In other examples, the methods may further comprise using thenucleic acids produced from plants, plant parts, plant cells or seeds ina plant breeding program, for example in making crosses, selectionand/or advancement decisions in a breeding program. Crossing includesany type of plant breeding crossing method, including but not limited tocrosses to produce hybrids, outcrossing, selfing, backcrossing, locusconversion, introgression and the like. Favorable genotypes and ormarker profiles, optionally associated with a trait of interest, may beidentified by one or more methodologies. In some examples one or moremarkers are used, including but not limited to restriction fragmentlength polymorphism (RFLP), random amplified polymorphic DNA (RAPD),amplified fragment length polymorphism (AFLP), inter-simple sequencerepeats (ISSRs), sequence characterized regions (SCARs), sequence tagsites (STSs), cleaved amplified polymorphic sequences (CAPS),microsatellites, simple sequence repeats (SSRs), expressed sequence tags(ESTs), single nucleotide polymorphisms (SNPs), and diversity arraystechnology (DArT), sequencing, and the like. In some methods, a targetnucleic acid is amplified prior to hybridization with a probe. In othercases, the target nucleic acid is not amplified prior to hybridization,such as methods using molecular inversion probes. In some examples, thegenotype related to a specific trait is monitored, while in otherexamples, a genome-wide evaluation including but not limited to one ormore of marker panels, library screens, association studies,microarrays, gene chips, expression studies, or sequencing such aswhole-genome resequencing and genotyping-by-sequencing (GBS) may beused. In some examples, no target-specific probe is needed, for exampleby using sequencing technologies, including but not limited tonext-generation sequencing methods (see, for example, Metzker (2010) NatRev Genet 11:31-46; and, Egan et al. (2012) Am J Bot 99:175-185) such assequencing by synthesis (e.g., Roche 454 pyrosequencing, Illumina GenomeAnalyzer, and Ion Torrent PGM or Proton systems), sequencing by ligation(e.g., SOLiD from Applied Biosystems, and Polnator system from AzcoBiotech), and single molecule sequencing (SMS or third-generationsequencing) which eliminate template amplification (e.g., Helicossystem, and PacBio RS system from Pacific BioSciences). Furthertechnologies include optical sequencing systems (e.g., Starlight fromLife Technologies), and nanopore sequencing (e.g., GridION from OxfordNanopore Technologies). Each of these may be coupled with one or moreenrichment strategies for organellar or nuclear genomes in order toreduce the complexity of the genome under investigation via PCR,hybridization, restriction enzyme (see, e.g., Elshire et al. (2011) PLoSONE 6:e19379), and expression methods. In some examples, no referencegenome sequence is needed in order to complete the analysis. N00655 andits plant parts can be identified through a molecular marker profile.Such plant parts may be either diploid or haploid. Also encompassed anddescribed are plants and plant parts substantially benefiting from theuse of variety N00655 in their development, such as variety N00655comprising a locus conversion or single locus conversion.

The production of doubled haploids can also be used for the developmentof inbreds from N00655 in a breeding program. In Brassica napus,microspore culture technique is used in producing haploid embryos. Thehaploid embryos are then regenerated on appropriate media as haploidplantlets, doubling chromosomes of which results in doubled haploidplants. This can be advantageous because the process omits thegenerations of selfing needed to obtain a homozygous plant from aheterozygous source.

Controlling Self-Pollination

Canola varieties are mainly self-pollinated. A pollination controlsystem and effective transfer of pollen from one parent to the otherprovides an effective method for producing hybrid canola seed andplants. For example, the ogura cytoplasmic male sterility (CMS) system,developed via protoplast fusion between radish (Raphanus sativus) andrapeseed (Brassica napus), is one of the most frequently used methods ofhybrid production. It provides stable expression of the male sterilitytrait and an effective nuclear restorer gene. The OGU INRA restorergene, Rf1 originating from radish has improved versions.

Brassica hybrid varieties can be developed using self-incompatible (SI),cytoplasmic male sterile (CMS) or nuclear male sterile (NMS) Brassicaplants as the female parent such that only cross pollination will occurbetween the hybrid parents.

In one instance, production of F₁ hybrids includes crossing a CMSBrassica female parent with a pollen-producing male Brassica has afertility restorer gene (Rf gene). The presence of an Rf gene means thatthe F₁ generation will not be completely or partially sterile, so thateither self-pollination or cross pollination may occur. Self pollinationof the F₁ generation to produce several subsequent generations verifiesthat a desired trait is heritable and stable and that a new variety hasbeen isolated.

Other sources and refinements of CMS sterility in canola include thePolima cytoplasmic male sterile plant, as well as those of U.S. Pat. No.5,789,566, DNA sequence imparting cytoplasmic male sterility,mitochondrial genome, nuclear genome, mitochondria and plant containingsaid sequence and process for the preparation of hybrids; See U.S. Pat.Nos. 4,658,085, 5,973,233 and 6,229,072.

Hybrid Development

For many traits the true genotypic value may be masked by otherconfounding plant traits or environmental factors. One method foridentifying a superior plant is to observe its performance relative toother experimental plants and to one or more widely grown standardvarieties. If a single observation is inconclusive, replicatedobservations provide a better estimate of the genetic worth.

As a result of the advances in sterility systems, lines are developedthat can be used as an open pollinated variety (i.e., a purelinecultivar) and/or as a sterile inbred (female) used in the production ofF₁ hybrid seed. In the latter case, favorable combining ability with arestorer (male) would be desirable.

The development of a canola hybrid generally involves three steps: (1)the selection of plants from various germplasm pools for initialbreeding crosses; (2) generation of inbred lines, such as by selfing ofselected plants from the breeding crosses for several generations toproduce a series of different inbred lines, which breed true and arehighly uniform; and (3) crossing the selected inbred lines withdifferent inbred lines to produce the hybrids.

Combining ability of a line, as well as the performance of the line perse, is a factor in the selection of improved canola lines that may beused as inbreds. Combining ability refers to a line's contribution as aparent when crossed with other lines to form hybrids. The hybrids formedfor the purpose of selecting superior lines are designated test crosses.One way of measuring combining ability is by using breeding values.Breeding values are based on the overall mean of a number of testcrosses. This mean is then adjusted to remove environmental effects andit is adjusted for known genetic relationships among the lines.

Brassica napus canola plants, absent the use of sterility systems, arerecognized to commonly be self-fertile with approximately 70 to 90percent of the seed normally forming as the result of self-pollination.The percentage of cross pollination may be further enhanced whenpopulations of recognized insect pollinators at a given growing site aregreater. Thus open pollination is often used in commercial canolaproduction.

Locus Conversions of Canola Variety N00655

N00655 represents a new base genetic line into which a new locus ortrait may be introduced. Direct transformation, genetic editing or genemodification such as described herein and backcrossing can be used toaccomplish such an introgression. The term locus conversion is used todesignate the product of such an introgression.

Backcrossing can be used to improve inbred varieties and a hybridvariety which is made using those inbreds. Backcrossing can be used totransfer a specific desirable trait from one variety, the donor parent,to an inbred called the recurrent parent which has overall goodagronomic characteristics yet that lacks the desirable trait. Thistransfer of the desirable trait into an inbred with overall goodagronomic characteristics can be accomplished by first crossing arecurrent parent to a donor parent (non-recurrent parent). The progenyof this cross is then mated back to the recurrent parent followed byselection in the resultant progeny for the desired trait to betransferred from the non-recurrent parent.

Traits may be used by those of ordinary skill in the art to characterizeprogeny. Traits are commonly evaluated at a significance level, such asa 1%, 5% or 10% significance level, when measured in plants grown in thesame environmental conditions. For example, a locus conversion of N00655may be characterized as having essentially the same phenotypic traits asN00655 or otherwise all of the physiological and morphologicalcharacteristics of N00655. The traits used for comparison may be thosetraits shown in Table 1. Molecular markers can also be used during thebreeding process for the selection of qualitative traits. For example,markers can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select for the genome of the recurrent parent and against thegenome of the donor parent. Using this procedure can minimize the amountof genome from the donor parent that remains in the selected plants.

A locus conversion of N00655 may contain at least 1, 2, 3, 4 or 5 locusconversions, and fewer than 15, 10, 9, 8, 7, or 6 locus conversions. Alocus conversion of N00655 will otherwise retain the genetic integrityof N00655. For example, a locus conversion of N00655 can be developedwhen DNA sequences are introduced through backcrossing, with a parent ofN00655 utilized as the recurrent parent. Both naturally occurring andtransgenic DNA sequences may be introduced through backcrossingtechniques. A backcross conversion may produce a plant with a locusconversion in at least one or more backcrosses, including at least 2crosses, at least 3 crosses, at least 4 crosses, at least 5 crosses andthe like. Molecular marker assisted breeding or selection may beutilized to reduce the number of backcrosses necessary to achieve thebackcross conversion. For example, a backcross conversion can be made inas few as two backcrosses. A locus conversion of N00655 can bedetermined through the use of a molecular profile. A locus conversion ofN00655 may have at least 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of themolecular markers, or molecular profile, of N00655. Examples ofmolecular markers that could be used to determine the molecular profileinclude RFLP, PCR analysis, SSR and SNPs.

Examples of locus conversions or transgenes which may be using includeone or more that confer male sterility, a site for site-specificrecombination, abiotic stress tolerance, altered phosphate content,altered antioxidants, altered fatty acid content, altered essentialamino acid content, altered carbohydrate content, herbicide resistance,insect resistance, disease resistance or a combination thereof. Otherdesirable traits which may be modified include tolerance to heat anddrought, reducing the time to crop maturity, greater yield, and betteragronomic quality, increased amount or rate of germination, standestablishment, growth rate, maturity, and plant and pod height.

Disease—Sclerotinia

Sclerotinia infects over 100 species of plants, including Brassicaspecies. Sclerotinia sclerotiorum is responsible for over 99% ofSclerotinia disease, while Sclerotinia minor produces less than 1% ofthe disease. Sclerotinia produces sclerotia, irregularly-shaped, darkoverwintering bodies, which can endure in soil for four to five years.The sclerotia can germinate carpogenically or myceliogenically,depending on the environmental conditions and crop canopies. The twotypes of germination cause two distinct types of diseases. Sclerotiathat germinate carpogenically produce apothecia and ascospores thatinfect above-ground tissues, resulting in stem blight, stalk rot, headrot, pod rot, white mold and blossom blight of plants. Sclerotia thatgerminate myceliogenically produce mycelia that infect root tissues,causing crown rot, root rot and basal stalk rot.

Sclerotinia causes Sclerotinia stem rot, also known as white mold, inBrassica, including canola. The disease is favored by moist soilconditions (at least 10 days at or near field capacity) and temperaturesof 15-25° C., prior to and during canola flowering. The spores cannotinfect leaves and stems directly; they must first land on flowers,fallen petals, and pollen on the stems and leaves. The fungal spores usethe flower parts as a food source as they germinate and infect theplant.

The severity of Sclerotinia in Brassica is variable, and is dependent onthe time of infection and climatic conditions, being favored by cooltemperatures between 20 and 25° C., prolonged precipitation and relativehumidities of greater than 80%. Losses ranging from 5 to 100% have beenreported for individual fields. Sclerotinia can cause heavy losses inwet swaths and result in economic losses of millions of dollars.

The symptoms of Sclerotinia infection usually develop several weeksafter flowering begins. The infections often develop where the leaf andthe stem join. Infected stems appear bleached and tend to shred. Hardblack fungal sclerotia develop within the infected stems, branches, orpods. Plants infected at flowering produce little or no seed. Plantswith girdled stems wilt and ripen prematurely. Severely infected cropsfrequently lodge, shatter at swathing, and make swathing more timeconsuming. Infections can occur in all above-ground plant parts,especially in dense or lodged stands, where plant-to-plant contactfacilitates the spread of infection. New sclerotia carry the diseaseover to the next season.

Conventional methods for control of Sclerotinia diseases include (a)chemical control (fungicides such as benomyl, vinclozolin, iprodione,azoxystrobin, prothioconazole, boscalid), (b) disease resistance (suchas partial resistance and breeding for favorable morphologies such asincreased standability, reduced petal retention, branching (less compactand/or higher), and early leaf abscission) and (c) cultural control.

Methods for generating Sclerotinia resistant Brassica plants usinginbred line N00655 are provided, including crossing with one or morelines containing one or more genes contributing to Sclerotiniaresistance and selecting for resistance. In some embodiments, N00655 canbe modified to have resistance to Sclerotinia.

The inbred line N00655 can be used in breeding techniques to createcanola hybrids. For example, inbred line N00655 may be used as a femaleparent, male parent or restorer (R-line), A-line, maintainer (B-line) ina canola hybrid.

An OGU restorer version, or R-line, of variety N00655 is provided whichis a male line that carries a gene for the restoration of fertility.When a sterile CMS version of an inbred is pollinated by a male linethat carries a gene for the restoration of fertility, it results in afertile hybrid. Generally, the seed produced from this cross is the seedthat is commercially sold.

There are a number of analytical methods available to determine thephenotypic stability of a canola variety. Phenotypic trait data areusually collected in field experiments including for example traitsassociated with seed yield, seed oil content, seed protein content,fatty acid composition of oil, glucosinolate content of meal, growthhabit, lodging resistance, plant height, shattering resistance, etc.

In addition to phenotypic observations, the genotype of a plant can alsobe examined. A plant's genotype can be used to identify plants of thesame variety or a related variety, or pedigree. Genotyping techniquesinclude Isozyme Electrophoresis, RFLPs, RAPDs, AP-PCR, DAF, SCARs,AFLPs, SSRs which are also referred to as Microsatellites and SNPs.

The variety described herein has shown uniformity and stability for alltraits, such as described in Table 1. When preparing the detailedphenotypic information, plants of variety N00655 were observed whilebeing grown using conventional agronomic practices.

Variety N00655 can be advantageously used in accordance with thebreeding methods described herein and those known in the art to producehybrids and other progeny plants retaining desired trait combinations ofN00655. Provided are methods for producing a canola plant by crossing afirst parent canola plant with a second parent canola plant whereineither the first or second parent canola plant is canola variety N00655.Further, both first and second parent canola plants can come from thecanola variety N00655. Either the first or the second parent plant maybe male sterile.

Still further, methods to produce a N00655-derived canola plant areprovided by crossing canola variety N00655 with a second canola plantand growing the progeny seed, and repeating the crossing and growingsteps with the canola N00655-derived plant at least 1, 2 or 3 times andless than 7, 6, 5, 4, 3 or 2 times. Any such methods using the canolavariety N00655 may include one or more of open pollination, selfing,backcrosses, hybrid production, crosses to populations, and the like.All plants produced using canola variety N00655 as a parent, includingplants derived from canola variety N00655 are provided herein. Plantsderived or produced from N00655 may include components for either malesterility or for restoration of fertility. Advantageously, the canolavariety is used in crosses with other, different, canola plants toproduce first generation (F₁) canola hybrid seeds and plants withsuperior characteristics.

A single-gene or a single locus conversion of N00655 is provided.Single-gene conversions and single locus conversions can occur when DNAsequences are introduced through traditional (non-transformation)breeding techniques, such as backcrossing. DNA sequences, whethernaturally occurring, modified or transgenes, may be introduced usingthese traditional breeding techniques. Desired traits transferredthrough this process include, but are not limited to, fertilityrestoration, fatty acid profile modification, oil content modification,protein quality or quantity modification, other nutritionalenhancements, industrial enhancements, disease resistance, insectresistance, herbicide resistance and yield enhancements. The trait ofinterest is transferred from the donor parent to the recurrent parent,in this case, the canola plant disclosed herein. Single-gene traits mayresult from the transfer of either a dominant allele or a recessiveallele. Selection of progeny containing the trait of interest is done bydirect selection for a trait associated with a dominant allele.Selection of progeny for a trait that is transferred via a recessiveallele will require growing and selfing the first backcross to determinewhich plants carry the recessive alleles. Recessive traits may requireadditional progeny testing in successive backcross generations todetermine the presence of the gene of interest.

It should be understood that the canola variety described herein can,through routine manipulation by cytoplasmic genes, nuclear genes, orother factors, be produced in a male-sterile or restorer form asdescribed in the references discussed earlier. Canola variety N00655 canbe manipulated to be male sterile by any of a number of methods known inthe art, including by the use of mechanical methods, chemical methods,SI, CMS (either ogura or another system) or NMS. The term “manipulatedto be male sterile” refers to the use of any available techniques toproduce a male sterile version of canola variety N00655. The malesterility may be either partial or complete male sterility. F1 hybridseed and plants produced by the use of canola variety N00655 areprovided. Canola variety N00655 can also further comprise a componentfor fertility restoration of a male sterile plant, such as an Rfrestorer gene. In this case, canola variety N00655 could then be used asthe male plant in hybrid seed production.

N00655 can be used in tissue culture. As used herein, the term plantincludes plant protoplasts, plant cell tissue cultures from which canolaplants can be regenerated, plant calli, plant clumps, and plant cellsthat are intact in plants or parts of plants, such as embryos, pollen,ovules, seeds, flowers, leaves, husks, stalks, roots, root tips, anthersand the like. Tissue culture and microspore cultures and theregeneration of canola plants therefrom are provided.

The utility of canola variety N00655 also extends to crosses with otherspecies than just Brassica napus. Commonly, suitable species will be ofthe family Brassicae.

Molecular biological techniques allow the isolation and characterizationof genetic elements with specific functions, such as encoding specificprotein products. The genome of plants can be engineered to contain andexpress foreign genetic elements, or additional or modified versions ofnative or endogenous genetic elements in order to alter the traits of aplant in a specific manner. Any DNA sequences, whether from a differentspecies or from the same species, that are inserted into the genomeusing transformation are referred to herein collectively as“transgenes”. Gene editing can insert, delete or substitute nativepolynucleotide sequences to produce increased or decreased expression oractivity of a polypeptide of interest. Described herein are transformedand edited versions of the claimed canola variety N00655.

Numerous methods for plant transformation have been developed, includingbiological and physical plant transformation protocols. In addition,expression vectors and in vitro culture methods for plant cell or tissuetransformation and regeneration of plants are available. The mostprevalent types of plant transformation involve the construction of anexpression vector. Such a vector comprises a DNA sequence that containsa gene under the control of or operatively linked to a regulatoryelement, for example a promoter. The vector may contain one or moregenes and one or more regulatory elements.

In general, methods to transform, modify, edit or alter plant endogenousgenomic DNA include altering the plant native DNA sequence orintroducing a pre-existing transgenic sequence including regulatoryelements, coding and non-coding sequences. Genetic transformationmethods include introduction of foreign or heterologous sequences andgenome editing techniques which modify the native sequence.Transformation methods can be used, for example, to target nucleic acidsto pre-engineered target recognition sequences in the genome. Suchpre-engineered target sequences may be introduced by genome editing ormodification. As an example, a genetically modified plant variety isgenerated using “custom” or engineered endonucleases such asmeganucleases produced to modify plant genomes (see e.g., WO2009/114321; Gao et al. (2010) Plant Journal 1:176-187). Anothersite-directed engineering method is through the use of zinc fingerdomain recognition coupled with the restriction properties ofrestriction enzyme. See e.g., Urnov, et al., (2010) Nat Rev Genet.11(9):636-46; Shukla, et al., (2009) Nature 459 (7245):437-41. Atranscription activator-like (TAL) effector-DNA modifying enzyme (TALEor TALEN) is also used to engineer changes in plant genome. See e.g.,US20110145940, Cermak et al., (2011) Nucleic Acids Res. 39(12) and Bochet al., (2009), Science 326(5959): 1509-12. Site-specific modificationof plant genomes can also be performed using the bacterial type IICRISPR (clustered regularly interspaced short palindromic repeats)/Cas(CRISPR-associated) system. See e.g., Belhaj et al., (2013), PlantMethods 9: 39; The Cas9/guide RNA-based system allows targeted cleavageof genomic DNA guided by a customizable small noncoding RNA in plants(see e.g., WO 2015026883A1).

A genetic trait which has been engineered into a particular canola plantusing transformation and/or gene editing techniques, could be moved intoanother line using traditional breeding techniques that are well knownin the plant breeding arts. For example, a backcrossing approach couldbe used to move a transgene or modified gene from a transformed ormodified canola plant to an elite inbred line and the resulting progenywould comprise a transgene or modified gene. Also, if an inbred line wasused for the transformation or genetic modification then the transgenicor modified plants could be crossed to a different line in order toproduce a transgenic or modified hybrid canola plant. As used herein,“crossing” can refer to a simple X by Y cross, or the process ofbackcrossing, depending on the context. Various genetic elements can beintroduced into the plant genome using transformation or gene editing.These elements include but are not limited to genes; coding sequences;inducible, constitutive, and tissue specific promoters; enhancingsequences; and signal and targeting sequences.

Transgenic and modified plants described herein can produce a foreign ormodified protein in commercial quantities. Thus, techniques for theselection and propagation of transformed plants, which are wellunderstood in the art, may yield a plurality of transgenic or modifiedplants which are harvested in a conventional manner, and a foreign ormodified protein then can be extracted from a tissue of interest or fromtotal biomass.

A genetic map can be generated, for example via conventional RFLP, PCRanalysis, SSR and SNPs, which identifies the approximate chromosomallocation of the integrated DNA molecule coding for the foreign protein.Genetic or physical map information concerning chromosomal location isuseful for proprietary protection of a subject transgenic or modifiedplant. If unauthorized propagation is undertaken and crosses made withother germplasm, the map of the integration or modified region can becompared to similar maps for suspect plants, to determine if the latterhave a common parentage with the subject plant. Map comparisons wouldinvolve hybridizations, RFLP, PCR, SSR, SNP, and sequencing, all ofwhich are conventional techniques.

Likewise, disclosed are plants genetically engineered or modified toexpress various phenotypes of agronomic interest. Exemplary transgenesor modified genes implicated in this regard include, but are not limitedto, those categorized below.

1. Genes that Confer Resistance to Pests or Disease and that Encode:

(A) Plant disease resistance genes. Plant defenses are often activatedby specific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with clonedresistance gene to engineer plants that are resistant to specificpathogen strains. A plant resistant to a disease is one that is moreresistant to a pathogen as compared to the wild type plant.

(B) A gene conferring resistance to fungal pathogens.

(C) A Bacillus thuringiensis protein, a derivative thereof or asynthetic polypeptide modeled thereon. DNA molecules encodingdelta-endotoxin genes can be purchased from American Type CultureCollection (Manassas, Va.), for example, under ATCC Accession Nos.40098, 67136, 31995 and 31998. Other examples of Bacillus thuringiensistransgenes are given in the following US and international patents andapplications: U.S. Pat. Nos. 5,188,960; 5,689,052; 5,880,275; WO91/114778; WO 99/31248; WO 01/12731; WO 99/24581; WO 97/40162.

(D) An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof.

(E) An insect-specific peptide which, upon expression, disrupts thephysiology of the affected pest. For example, DNA coding for insectdiuretic hormone receptor, allostatins and genes encodinginsect-specific, paralytic neurotoxins.

(F) An enzyme responsible for a hyperaccumulation of a monterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

(G) An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTApplication No. WO 93/02197, which discloses the nucleotide sequence ofa callase gene. DNA molecules which contain chitinase-encoding sequencescan be obtained, for example, from the ATCC under Accession Nos. 39637and 67152. See also U.S. Pat. No. 6,563,020.

(H) A molecule that stimulates signal transduction. For example,nucleotide sequences encoding calmodulin.

(I) A hydrophobic moment peptide. See, U.S. Pat. Nos. 5,580,852 and5,607,914.

(J) A membrane permease, a channel former or a channel blocker. Forexample, a cecropin-beta lytic peptide analog.

(K) A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. Coat protein-mediated resistance has beenconferred upon transformed plants against alfalfa mosaic virus, cucumbermosaic virus, tobacco streak virus, potato virus X, potato virus Y,tobacco etch virus, tobacco rattle virus and tobacco mosaic virus.

(L) An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect.

(M) A virus-specific antibody. For example, transgenic plants expressingrecombinant antibody genes can be protected from virus attack.

(N) A developmental-arrestive protein produced in nature by a pathogenor a parasite; for example, fungal endo alpha-1,4-D-polygalacturonases.

(O) A developmental-arrestive protein produced in nature by a plant.

(P) Genes involved in the Systemic Acquired Resistance (SAR) Responseand/or the pathogenesis related genes.

(Q) Antifungal genes.

(R) Detoxification genes, such as for fumonisin, beauvericin,moniliformin and zearalenone and their structurally related derivatives.For example, see, U.S. Pat. No. 5,792,931.

(S) Cystatin and cysteine proteinase inhibitors. E.g., U.S. Pat. No.7,205,453.

(T) Defensin genes.

(U) Genes that confer resistance to Phytophthora Root Rot, such as theBrassica equivalents of the Rps 1, Rps 1-a, Rps 1-b, Rps 1-c, Rps 1-d,Rps 1-e, Rps 1-k, Rps 2, Rps 3-a, Rps 3-b, Rps 3-c, Rps 4, Rps 5, Rps 6,Rps 7 and other Rps genes.

2. Genes that Confer Resistance to an Herbicide, for Example:

(A) A herbicide that inhibits the growing point or meristem, such as animidazalinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme. See also, U.S. Pat. Nos. 5,605,011;5,013,659; 5,141,870; 5,767,361; 5,731,180; 5,304,732; 4,761,373;5,331,107; 5,928,937 and 5,378,824.

(B) Glyphosate (resistance imparted by mutant5-enolpyruvl-3-phosphikimate synthase (EPSP) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase, PAT) and Streptomyceshygroscopicus phosphinothricin-acetyl transferase, bar, genes), andpyridinoxy or phenoxy propionic acids and cycloshexones (ACCaseinhibitor-encoding genes). See, for example, U.S. Pat. No. 4,940,835,which discloses the nucleotide sequence of a form of EPSP which canconfer glyphosate resistance. See also, U.S. Pat. No. 7,405,074, andrelated applications, which disclose compositions and means forproviding glyphosate resistance. U.S. Pat. No. 5,627,061 describes genesencoding EPSPS enzymes. See also, U.S. Pat. Nos. 6,566,587; 6,338,961;6,248,876; 6,040,497; 5,804,425; 5,633,435; 5,145,783; 4,971,908;5,312,910; 5,188,642; 4,940,835; 5,866,775; 6,225,114; 6,130,366;5,310,667; 4,535,060; 4,769,061; 5,633,448; 5,510,471; Re. 36,449; RE37,287 E; and 5,491,288; and international publications EP1173580; WO01/66704; EP1173581 and EP1173582. A DNA molecule encoding a mutant aroAgene can be obtained under ATCC Accession No. 39256, see U.S. Pat. No.4,769,061. European Patent Application No. 0 333 033, and U.S. Pat. No.4,975,374 disclose nucleotide sequences of glutamine synthetase geneswhich confer resistance to herbicides such as L-phosphinothricin. Thenucleotide sequence of a phosphinothricin-acetyl-transferase gene isprovided in European Application No. 0 242 246. See also, U.S. Pat. Nos.5,969,213; 5,489,520; 5,550,318; 5,874,265; 5,919,675; 5,561,236;5,648,477; 5,646,024; 6,177,616 and 5,879,903. Exemplary of genesconferring resistance to phenoxy propionic acids and cycloshexones, suchas sethoxydim and haloxyfop, are the Acc1-S1, Acc1-S2 and Acc1-S3 genes.See also, U.S. Pat. Nos. 5,188,642; 5,352,605; 5,530,196; 5,633,435;5,717,084; 5,728,925; 5,804,425 and Canadian Patent No. 1,313,830.

(C) A herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+ genes) and a benzonitrile (nitrilase gene). Nucleotide sequencesfor nitrilase genes are disclosed in U.S. Pat. No. 4,810,648, and DNAmolecules containing these genes are available under ATCC Accession Nos.53435, 67441 and 67442.

(D) Acetohydroxy acid synthase, which has been found to make plants thatexpress this enzyme resistant to multiple types of herbicides, has beenintroduced into a variety of plants. Other genes that confer toleranceto herbicides include: a gene encoding a chimeric protein of ratcytochrome P4507A1 and yeast NADPH-cytochrome P450 oxidoreductase, genesfor glutathione reductase and superoxide dismutase, and genes forvarious phosphotransferases.

(E) Protoporphyrinogen oxidase (protox) is necessary for the productionof chlorophyll, which is necessary for all plant survival. The protoxenzyme serves as the target for a variety of herbicidal compounds. Theseherbicides also inhibit growth of all the different species of plantspresent, causing their total destruction. The development of plantscontaining altered protox activity which are resistant to theseherbicides are described in U.S. Pat. Nos. 6,288,306; 6,282,837; and5,767,373; and international publication WO 01/12825.

3. Transgenes that Confer or Contribute to an Altered GrainCharacteristic, Such as:

(A) Altered fatty acids, for example, by

-   -   (1) Down-regulation of stearoyl-ACP desaturase to increase        stearic acid content of the plant. See, WO99/64579,    -   (2) Elevating oleic acid via FAD-2 gene modification and/or        decreasing linolenic acid via FAD-3 gene modification, See, U.S.        Pat. Nos. 6,063,947; 6,323,392; 6,372,965 and WO 93/11245,    -   (3) Altering conjugated linolenic or linoleic acid content, such        as in WO 01/12800,    -   (4) Altering LEC1, AGP, Dek1, Superal1, mi1ps, various Ipa genes        such as Ipa1, Ipa3, hpt or hggt. For example, see WO 02/42424,        WO 98/22604, WO 03/011015, U.S. Pat. Nos. 6,423,886, 6,197,561,        6,825,397, US Patent Application Publication Nos. 2003/0079247,        2003/0204870, WO02/057439, WO03/011015.

(B) Altered phosphorus content, for example, by the

-   -   (1) Introduction of a phytase-encoding gene would enhance        breakdown of phytate, adding more free phosphate to the        transformed plant, such as for example, using an Aspergillus        niger phytase gene.    -   (2) Up-regulation of a gene that reduces phytate content.

(C) Altered carbohydrates effected, for example, by altering a gene foran enzyme that affects the branching pattern of starch, a gene alteringthioredoxin. (See, U.S. Pat. No. 6,531,648). Exemplary genes includethose encoding fructosyltransferase, levansucrase, alpha-amylase,invertase, branching enzyme II, UDP-D-xylose 4-epimerase, Fragile 1 and2, Ref1, HCHL (4-hydroxycinnamoyl-CoA hydratase/lyase), C4H (cinnamate4-hydroxylase), AGP (ADPglucose pyrophosphorylase). The fatty acidmodification genes may also be used to affect starch content and/orcomposition through the interrelationship of the starch and oilpathways.

(D) Altered antioxidant content or composition, such as alteration oftocopherol or tocotrienols. For example, see, U.S. Pat. No. 6,787,683,US Patent Application Publication No. 2004/0034886 and WO 00/68393involving the manipulation of antioxidant levels through alteration of aphytl prenyl transferase (ppt), WO 03/082899 through alteration of ahomogentisate geranyl geranyl transferase (hggt).

(E) Altered essential seed amino acids. For example, see, U.S. Pat. No.6,127,600 (method of increasing accumulation of essential amino acids inseeds), U.S. Pat. No. 6,080,913 (binary methods of increasingaccumulation of essential amino acids in seeds), U.S. Pat. No. 5,990,389(high lysine), WO99/40209 (alteration of amino acid compositions inseeds), WO99/29882 (methods for altering amino acid content ofproteins), U.S. Pat. No. 5,850,016 (alteration of amino acidcompositions in seeds), WO98/20133 (proteins with enhanced levels ofessential amino acids), U.S. Pat. No. 5,885,802 (high methionine), U.S.Pat. No. 5,885,801 (high threonine), U.S. Pat. No. 6,664,445 (plantamino acid biosynthetic enzymes), U.S. Pat. No. 6,459,019 (increasedlysine and threonine), U.S. Pat. No. 6,441,274 (plant tryptophansynthase beta subunit), U.S. Pat. No. 6,346,403 (methionine metabolicenzymes), U.S. Pat. No. 5,939,599 (high sulfur), U.S. Pat. No. 5,912,414(increased methionine), WO98/56935 (plant amino acid biosyntheticenzymes), WO98/45458 (engineered seed protein having higher percentageof essential amino acids), WO98/42831 (increased lysine), U.S. Pat. No.5,633,436 (increasing sulfur amino acid content), U.S. Pat. No.5,559,223 (synthetic storage proteins with defined structure containingprogrammable levels of essential amino acids for improvement of thenutritional value of plants), WO96/01905 (increased threonine),WO95/15392 (increased lysine), US Patent Application Publication No.2003/0163838, US Patent Application Publication No. 2003/0150014, USPatent Application Publication No. 2004/0068767, U.S. Pat. No.6,803,498, WO01/79516, and WO00/09706 (Ces A: cellulose synthase), U.S.Pat. No. 6,194,638 (hemicellulose), U.S. Pat. No. 6,399,859 and USPatent Application Publication No. 2004/0025203 (UDPGdH), U.S. Pat. No.6,194,638 (RGP).

4. Genes that Control Pollination, Hybrid Seed Production orMale-Sterility:

There are several methods of conferring genetic male sterilityavailable, such as multiple mutant genes at separate locations withinthe genome that confer male sterility, as disclosed in U.S. Pat. Nos.4,654,465 and 4,727,219 and chromosomal translocations, see U.S. Pat.Nos. 3,861,709 and 3,710,511. U.S. Pat. No. 5,432,068 describes a systemof nuclear male sterility which includes replacing the native promoterof an essential male fertility gene with an inducible promoter to createa male sterile plant that can have fertility restored by inducing orturning “on”, the promoter such that the male fertility gene istranscribed.

(A) Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN-Ac-PPT (WO 01/29237).

(B) Introduction of various stamen-specific promoters (WO 92/13956, WO92/13957).

(C) Introduction of the barnase and the barstar gene.

For additional examples of nuclear male and female sterility systems andgenes, see also, U.S. Pat. Nos. 5,859,341; 6,297,426; 5,478,369;5,824,524; 5,850,014 and 6,265,640.

Also see, U.S. Pat. No. 5,426,041 (relating to a method for thepreparation of a seed of a plant comprising crossing a male sterileplant and a second plant which is male fertile), U.S. Pat. No. 6,013,859(molecular methods of hybrid seed production) and U.S. Pat. No.6,037,523 (use of male tissue-preferred regulatory region in mediatingfertility).

5. Genes that Create a Site for Site Specific DNA Integration.

This includes the introduction of FRT sites that may be used in theFLP/FRT system and/or Lox sites that may be used in the Cre/Loxp system.Other systems that may be used include the Gin recombinase of phage Mu,the Pin recombinase of E. coli, and the R/RS system of the pSR1 plasmid.

6. Genes that affect abiotic stress resistance (including but notlimited to flowering and seed development, enhancement of nitrogenutilization efficiency, altered nitrogen responsiveness, droughtresistance or tolerance, cold resistance or tolerance, and saltresistance or tolerance) and increased yield under stress.

For example, see, U.S. Pat. Nos. 5,892,009, 5,965,705, 5,929,305,5,891,859, 6,417,428, 6,664,446, 6,706,866, 6,717,034, 6,801,104. CBFgenes and transcription factors effective in mitigating the negativeeffects of freezing, high salinity, and drought on plants can be used.Altering abscisic acid in plants may result in increased yield and/orincreased tolerance to abiotic stress. Modifying cytokinin expressionmay result in plants with increased drought tolerance, and/or increasedyield. Enhancement of nitrogen utilization and altered nitrogenresponsiveness can be carried out. Ethylene alteration, planttranscription factors or transcriptional regulators of abiotic stressmay be used. Other genes and transcription factors that affect plantgrowth and agronomic traits such as yield, flowering, plant growthand/or plant structure, can be introduced or introgressed into plants.

Seed Treatments and Cleaning

Methods of harvesting the seed of the canola variety N00655 as seed forplanting are provided. Embodiments include cleaning the seed, treatingthe seed, and/or conditioning the seed. Cleaning the seed is understoodin the art to include removal of foreign debris such as one or more ofweed seed, chaff, and plant matter, from the seed. Conditioning the seedis understood in the art to include controlling the temperature and rateof dry down of the seed and storing seed in a controlled temperatureenvironment. Seed treatment is the application of a composition to thesurface of the seed such as a coating or powder. Methods for producing atreated seed include the step of applying a composition to the seed orseed surface. Seeds are provided which have on the surface acomposition. Biological active components such as bacteria can also beused as a seed treatment. Some examples of compositions areinsecticides, fungicides, pesticides, antimicrobials, germinationinhibitors, germination promoters, cytokinins, and nutrients.

Seed material can be treated, typically surface treated, with acomposition comprising combinations of chemical or biologicalherbicides, herbicide safeners, insecticides, fungicides, germinationinhibitors and enhancers, nutrients, plant growth regulators andactivators, bactericides, nematicides, avicides and/or molluscicides.These compounds are typically formulated together with further carriers,surfactants or application-promoting adjuvants customarily employed inthe art of formulation. The coatings may be applied by impregnatingpropagation material with a liquid formulation or by coating with acombined wet or dry formulation.

Some seed treatments that may be used on crop seed include, but are notlimited to, one or more of abscisic acid, acibenzolar-S-methyl,avermectin, amitrol, azaconazole, azospirillum, azadirachtin,azoxystrobin, Bacillus spp. (including one or more of cereus, firmus,megaterium, pumilis, sphaericus, subtilis and/or thuringiensis),Bradyrhizobium spp. (including one or more of betae, canariense,elkanii, iriomotense, japonicum, liaonigense, pachyrhizi and/oryuanmingense), captan, carboxin, chitosan, clothianidin, copper,cyazypyr, difenoconazole, etidiazole, fipronil, fludioxonil,fluoxastrobin, fluquinconazole, flurazole, fluxofenim, harpin protein,imazalil, imidacloprid, ipconazole, isoflavenoids,lipo-chitooligosaccharide, mancozeb, manganese, maneb, mefenoxam,metalaxyl, metconazole, myclobutanil, PCNB (EPA registration number00293500419, containing quintozen and terrazole), penflufen,penicillium, penthiopyrad, permethrine, picoxystrobin, prothioconazole,pyraclostrobin, rynaxypyr, S-metolachlor, saponin, sedaxane, TCMTB(2-(thiocyanomethylthio) benzothiazole), tebuconazole, thiabendazole,thiamethoxam, thiocarb, thiram, tolclofos-methyl, triadimenol,trichoderma, trifloxystrobin, triticonazole and/or zinc.

INDUSTRIAL APPLICABILITY

The seed of the N00655 variety or grain produced on its hybrids, plantsproduced from such seed, and various parts of the N00655 variety canolaplant or its progeny can be utilized in the production of an ediblevegetable oil, meal, other food products or silage for animal feed inaccordance with known techniques. The oil as removed from the seeds canbe used in food applications such as a salad or frying oil. Canola oilhas low levels of saturated fatty acids. “Canola” refers to rapeseed(Brassica) which (1) has an erucic acid (C22:1) content of at most 2%(preferably at most 0.5% or 0%) by weight based on the total fatty acidcontent of a seed, and (2) produces, after crushing, an air-dried mealcontaining less than 30 μmol glucosinolates per gram of defatted(oil-free) meal. The oil also finds utility in industrial applications.The solid meal component derived from seeds after oil extraction can beused as a nutritious livestock feed. Examples of canola grain as acommodity plant product include, but are not limited to, oils and fats,meals and protein, and carbohydrates. Methods of processing seeds andgrain of N00655 or of a hybrid and grain produced on the hybrid toproduce commodity products such as oil and protein meal are provided.

All publications, patents, and patent applications mentioned in thespecification are incorporated by reference herein for the purpose citedto the same extent as if each was specifically and individuallyindicated to be incorporated by reference herein.

The foregoing invention has been described in detail by way ofillustration and example for purposes of clarity and understanding. Asis readily apparent to one skilled in the art, the foregoing are onlysome of the methods and compositions that illustrate the embodiments ofthe foregoing invention. It will be apparent to those of ordinary skillin the art that variations, changes, modifications, and alterations maybe applied to the compositions and/or methods described herein withoutdeparting from the true spirit, concept, and scope of the invention.

As used herein, the terms “comprises,” “comprising,” “includes,”“including,” “has,” “having,” “contains”, “containing,” “characterizedby” or any other variation thereof, are intended to cover anon-exclusive inclusion.

Unless expressly stated to the contrary, “or” is used as an inclusiveterm. For example, a condition A or B is satisfied by any one of thefollowing: A is true (or present) and B is false (or not present), A isfalse (or not present) and B is true (or present), and both A and B aretrue (or present). The indefinite articles “a” and “an” preceding anelement or component are nonrestrictive regarding the number ofinstances (i.e., occurrences) of the element or component. Therefore “a”or “an” should be read to include one or at least one, and the singularword form of the element or component also includes the plural unlessthe number is obviously meant to be singular.

Deposits

Applicant has made a deposit of at least 625 seeds of canola line N00655with the American Type Culture Collection (ATCC), Manassas, Va. 20110USA, ATCC Deposit Number PTA-126287. The seeds deposited with the ATCCon Oct. 15, 2019 were taken from the deposit maintained by PioneerHi-Bred International, Inc., 7100 NW 62^(nd) Avenue, Johnston, Iowa50131-1000 since prior to the filing date of this application. Duringthe pendency of the application, access to this deposit will beavailable to the Commissioner of Patents and Trademarks and personsdetermined by the Commissioner to be entitled thereto upon request. Uponissuance of any claims in the application, the Applicant will make thedeposit available to the public, pursuant to 37 CFR 1.808. This depositof Canola line N00655 will be maintained in the ATCC depository, whichis a public depository, for a period of 30 years, or 5 years after themost recent request, or for the enforceable life of the patent,whichever is longer, and will be replaced if it becomes nonviable duringthat period. Additionally, Applicant(s) have or will satisfy all therequirements of 37 C.F.R. §§ 1.801-1.809, including providing anindication of the viability of the sample. Applicant(s) have noauthority to waive any restrictions imposed by law on the transfer ofbiological material or its transportation in commerce. Applicant(s) donot waive any infringement of their rights granted under this patent orunder the Plant Variety Protection Act (7 USC 2321 et seq.).

Breeding History

N00655 was developed from a complex cross of NS6227 (U.S. Pat. No.8,878,011) crossed with the result of the complex cross of [NS5870MC(U.S. Pat. No. 8,829,283) by a proprietary inbred] with [a differentproprietary inbred by ECD 04 (a publically available Brassica rapaline)]. The last crossing was completed and used to create a doubledhaploid population. The double haploid lines were evaluated for generalvigor, uniformity, maturity, oil, protein, total glucosinolates. Doublehaploids were tested for the level of resistance to clubroot. Selecteddouble haploid lines were evaluated in testcross hybrid seed productiontrials at six locations. Based on testcross hybrid data, N00655 wasselected.

TABLE 1 Table 1. Variety Descriptions based on Morphological, Agronomicand Quality Traits Morphological and Other Characteristics of CanolaBrassica napus N00655 Characteristic Value BEFORE FLOWERING CotyledonWidth (3 = Narrow, 5 = Medium, 7 = Wide) 5 Seedling Growth Habit (1 =Weak Rosette, 9 = Strong Rosette) 5 Stem Anthocyanin Intensity (1 =Absent or Very Weak, 3 = Weak, 5 = Medium, 7 = Strong, 9 = Very 1Strong) Leaf Type (1 = Petiolate, 9 = Lyrate) 3 Leaf Shape (3 = Narrowelliptic, 5 = Wide elliptic, 7 = Orbicular) Leaf Length (3 = Short, 5 =Medium, 7 = Long) 5 Leaf Width (3 = Narrow, 5 = Medium, 7 = Wide) 4 LeafColor (at 5-leaf stage) (1 = Light Green, 2 = Medium Green, 3 = DarkGreen, 4 = Blue-Green) 2 Leaf Waxiness (1 = Absent or Very Weak, 3 =Weak, 5 = Medium, 7 = Strong, 9 = Very Strong) Leaf Texture (1 = Smooth,9 = Rough) Leaf Lobe Development (1 = Absent or Very Weak, 3 = Weak, 5 =Medium, 7 = Strong, 9 = Very Strong) 3 Leaf Lobe Number (count) 2 LeafLobe Shape (1 = Acute, 9 = Rounded) Petiole Length (lobed cultivarsonly) (3 = Short, 5 = Medium, 7 = Long) 2 Leaf Margin Shape (1 =Undulating, 2 = Rounded, 3 = Sharp) 3 Leaf Margin Indentation (observefully developed upper stem leaves) (1 = Absent or Very Weak (very 4shallow), 3 = Weak (shallow), 5 = Medium, 7 = Strong (deep), 9 = VeryStrong (very deep)) Leaf Attachment to Stem (1 = Complete Clasping, 2 =Partial Clasping, 3 = Non-Clasping) 2 AFTER FLOWERING Time to Flowering(days from planting to 50% of plants showing one or more open flowers)47 Plant Height at Maturity (3 = Short, 5 = Medium, 7 = Tall) PlantGrowth Habit (1 = Erect, 3 = Semi-Erect, 5 = Intermediate, 7 =Semi-Prostrate, 9 = Prostrate) Flower-Bud Location (1 = Buds above mostrecently opened flowers, 9 = Buds below most recently opened 1 flowers)Petal Color (on first day of flowering) (1 = White, 2 = Light Yellow, 3= Medium Yellow, 4 = Dark Yellow, 3 5 = Orange, 6 = Other) Petal Length(3 = Short, 5 = Medium, 7 = Long) 5 Petal Width (3 = Narrow, 5 = Medium,7 = Wide) 5 Petal Spacing (1 = Open, 3 = Not Touching, 5 = Touching, 7 =Slight Overlap, 9 = Strong Overlap) 5 Anther Dotting (percentage atopening of flower) (1 = absent, 9 = present (percentage)) AntherArrangement (observe fully open flower) (1 = Introrse (facing inward), 2= Erect, 3 = Extrorse (facing outward)) Pod (silique) Length (1 = Short(<7 cm), 5 = Medium (7 to 10 cm), 9 = Long (>10 cm)) 4 Pod (silique)Width (3 = Narrow (3 mm), 5 = Medium (4 mm), 7 = Wide (5 mm)) 5 Pod(silique) Angle (1 = Erect, 3 = Semi-Erect, 5 = Horizontal, 7 = SlightlyDrooping, 9 = Drooping) 1 Pod (silique) Beak Length (3 = Short, 5 =Medium, 7 = Long) 5 Pod Pedicel Length (3 = Short, 5 = Medium, 7 = Long)5 Time to Maturity (days from planting to physiological maturity) 75SEED Seed Coat Color (1 = Black, 2 = Brown, 3 = Tan, 4 = Yellow, 5 =Mixed (describe), 6 = Other 1.5 (specify)) Seed Coat Mucilage (1 =absent, 9 = present) Seed Weight (5%-6% moisture) (grams per 1,000seeds) GRAIN QUALITY Oil Content (percentage, whole dry seed basis)45.91 Fatty-Acid Composition (percentage of total fatty acids in seedoil) 6.42 Palmitic Acid (C16:0) Stearic acid (C18:0) Oleic Acid (C18:1)Linoleic Acid (C18:2) Linolenic Acid (C18:3) Erucic Acid (C22:1) TotalSaturated Fats Protein Content (percentage in oil-free meal) 48.24Protein Content (percentage in whole dried seed) 26.09 Cystine CystosineMethionine Other (specify) Glucosinolate Content (μmoles of totalglucosinolates per gram whole seed, 8.5% moisture) (1 = Very 13.92 Low(<10 μmol per gram), 2 = Low (10-15 μmol per gram), 3 = Medium (15-20μmol per gram), 4 = High (>20 μmol per gram)) Chlorophyll Content (mg/kgseed, 8.5% moisture) (1 = Low (<8 ppm), 2 = Medium (8-15 ppm), 3 = High(>15 ppm))

What is claimed:
 1. A seed, plant, plant part or plant cell of canolavariety N00655, representative seed of said variety having beendeposited under ATCC accession number PTA-126287.
 2. The seed, plant,plant part, or plant cell of claim 1, wherein the seed, plant, plantpart, or plant cell is a plant part and wherein the plant part is anovule or pollen.
 3. An F1 hybrid Brassica seed of N00655 produced fromthe cross of the plant or plant part of claim 1 with a differentBrassica plant.
 4. An F1 hybrid Brassica plant or a plant part thereofproduced by growing the canola seed of claim 3, wherein the plant partcomprises at least one cell of the F1 hybrid Brassica plant.
 5. A methodfor producing a second Brassica plant, the method comprising applyingplant breeding techniques to the F1 plant or plant part of claim 4 toproduce the second Brassica plant.
 6. A method for producing a progenyBrassica seed, the method comprising (a) crossing an inducer varietywith the plant or plant part of claim 4 or a plant produced therefrom toproduce haploid seed, and (b) doubling the haploid seed therebyproducing the progeny Brassica seed.
 7. A method of making a commodityplant product comprising silage, carbohydrate, oil or protein, themethod comprising producing the commodity plant product from theBrassica plant or plant part of claim
 4. 8. A method of producing aBrassica seed derived from the variety N00655, the method comprising: a)crossing the plant of claim 1 with itself or a second plant to produceprogeny seed; and b) growing the progeny seed to produce a progeny plantand crossing the progeny plant with itself or a different plant toproduce Brassica seed derived from the variety N00655.
 9. A method forproducing nucleic acids, the method comprising isolating nucleic acidsfrom the seed, plant, plant part, or plant cell of claim
 1. 10. Aconverted seed, plant, plant part or plant cell of inbred canola varietyN00655, representative seed of the canola variety N00655 having beendeposited under ATCC accession number PTA-126287, wherein the convertedseed, plant, plant part or plant cell comprises a locus conversion, andwherein the plant or a plant grown from the converted seed, plant partor plant cell comprises the locus conversion and otherwise comprises thephysiological and morphological characteristics of canola variety N00655when grown under the same environmental conditions.
 11. The convertedseed, plant, plant part or plant cell of claim 10, wherein the locusconversion confers a property selected from the group consisting of malesterility, site-specific recombination, abiotic stress tolerance,altered phosphate, altered antioxidants, altered fatty acids, alteredessential amino acids, altered carbohydrates, herbicide tolerance,insect resistance and disease resistance.
 12. A Brassica seed producedby crossing the plant or plant part of claim 10 with a differentBrassica plant.
 13. A hybrid Brassica plant or plant part produced bygrowing the seed of claim 12, wherein the plant part comprises at leastone cell of the hybrid Brassica plant.
 14. A method for producing asecond Brassica plant, the method comprising applying plant breedingtechniques to the plant or plant part of claim 13 to produce the secondBrassica plant.
 15. A method for producing a second Brassica plant orplant part, the method comprising doubling haploid seed generated from across of the plant or plant part of claim 13 with an inducer variety,thereby producing the second Brassica plant or plant part.
 16. A methodof making a commodity plant product comprising carbohydrate, silage, oilor protein, the method comprising producing the commodity plant productfrom the Brassica plant or plant part of claim
 13. 17. A method forproducing nucleic acids, the method comprising isolating nucleic acidsfrom the seed, plant, plant part, or plant cell of claim
 10. 18. An F1hybrid seed produced by crossing a plant or plant part of inbred canolavariety N00655, representative seed of the variety having been depositedunder ATCC accession number PTA-126287 with a different Brassica plant,wherein inbred canola variety N00655 further comprises a transgene thatis inherited by the seed, wherein the transgene was introduced intoinbred canola variety N00655 by backcrossing or genetic transformation.19. A method of producing progeny seed, the method comprising crossing aplant grown from the seed of claim 18 with itself or a second plant toproduce progeny seed.
 20. A method of making a commodity plant productcomprising silage, carbohydrate, oil or protein, the method comprisingproducing the commodity plant product from an F1 plant grown from theseed of claim 18.